scholarly journals Proteomic Analyses of Vitreous in Proliferative Diabetic Retinopathy: Prior Studies and Future Outlook

2021 ◽  
Vol 10 (11) ◽  
pp. 2309
Author(s):  
Sarah R. Weber ◽  
Yuanjun Zhao ◽  
Christopher Gates ◽  
Jingqun Ma ◽  
Felipe da Veiga Leprevost ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Diabetic Retinopathy ◽  
Proliferative Diabetic Retinopathy ◽  
Proteomic Analysis ◽  
Retinal Disease ◽  
Vitreous Fluid ◽  
Future Outlook ◽  
Novel Proteins ◽  
Popular Medium ◽  
Molecular Information

Vitreous fluid is becoming an increasingly popular medium for the study of retinal disease. Numerous studies have demonstrated that proteomic analysis of the vitreous from patients with proliferative diabetic retinopathy yields valuable molecular information regarding known and novel proteins and pathways involved in this disease. However, there is no standardized methodology for vitreous proteomic studies. Here, we share a suggested protocol for such studies and outline the various experimental and analytic methods that are currently available. We also review prior mass spectrometry-based proteomic studies of the vitreous from patients with proliferative diabetic retinopathy, discuss common pitfalls of these studies, and propose next steps for moving the field forward.

scholarly journals A validated analysis pipeline for mass spectrometry-based vitreous proteomics: new insights into proliferative diabetic retinopathy

2021 ◽  
Vol 18 (1) ◽  
Author(s):  
Sarah R. Weber ◽  
Yuanjun Zhao ◽  
Jingqun Ma ◽  
Christopher Gates ◽  
Felipe da Veiga Leprevost ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Diabetic Retinopathy ◽  
Proliferative Diabetic Retinopathy ◽  
Metabolic Pathways ◽  
Power Analysis ◽  
Principal Component ◽  
Retinal Disease ◽  
Data Matrix ◽  
Accessible Information ◽  
Pooled Samples

Abstract Background Vitreous is an accessible, information-rich biofluid that has recently been studied as a source of retinal disease-related proteins and pathways. However, the number of samples required to confidently identify perturbed pathways remains unknown. In order to confidently identify these pathways, power analysis must be performed to determine the number of samples required, and sample preparation and analysis must be rigorously defined. Methods Control (n = 27) and proliferative diabetic retinopathy (n = 23) vitreous samples were treated as biologically distinct individuals or pooled together and aliquoted into technical replicates. Quantitative mass spectrometry with tandem mass tag labeling was used to identify proteins in individual or pooled control samples to determine technical and biological variability. To determine effect size and perform power analysis, control and proliferative diabetic retinopathy samples were analyzed across four 10-plexes. Pooled samples were used to normalize the data across plexes and generate a single data matrix for downstream analysis. Results The total number of unique proteins identified was 1152 in experiment 1, 989 of which were measured in all samples. In experiment 2, 1191 proteins were identified, 727 of which were measured across all samples in all plexes. Data are available via ProteomeXchange with identifier PXD025986. Spearman correlations of protein abundance estimations revealed minimal technical (0.99–1.00) and biological (0.94–0.98) variability. Each plex contained two unique pooled samples: one for normalizing across each 10-plex, and one to internally validate the normalization algorithm. Spearman correlation of the validation pool following normalization was 0.86–0.90. Principal component analysis revealed stratification of samples by disease and not by plex. Subsequent differential expression and pathway analyses demonstrated significant activation of metabolic pathways and inhibition of neuroprotective pathways in proliferative diabetic retinopathy samples relative to controls. Conclusions This study demonstrates a feasible, rigorous, and scalable method that can be applied to future proteomic studies of vitreous and identifies previously unrecognized metabolic pathways that advance understanding of diabetic retinopathy.

A Validated Analysis Pipeline for Mass Spectrometry-Based Vitreous Proteomics: Insights Into Proliferative Diabetic Retinopathy

2021 ◽  
Author(s):  
Sarah R Weber ◽  
Yuanjun Zhao ◽  
Jingqun Ma ◽  
Christopher Gates ◽  
Felipe da Veiga Leprevost ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Diabetic Retinopathy ◽  
Proliferative Diabetic Retinopathy ◽  
Power Analysis ◽  
Principal Component ◽  
Retinal Disease ◽  
Data Matrix ◽  
Accessible Information ◽  
Downstream Analysis ◽  
Pooled Samples

Abstract Background: Vitreous is an accessible, information-rich biofluid that has recently been studied as a source of retinal disease-related proteins and pathways. However, the number of samples required to confidently identify perturbed pathways remains unknown. In order to confidently identify these pathways, power analysis must be performed to determine the number of samples required.Methods: Control (n=27) and proliferative diabetic retinopathy (n=23) vitreous samples were treated as biologically distinct individuals or pooled together and aliquoted into technical replicates. Quantitative mass spectrometry with tandem mass tag labeling was used to identify proteins in individual or pooled control samples to determine technical and biological variability. To determine effect size and perform power analysis, control and proliferative diabetic retinopathy samples were analyzed across four 10plexes. Pooled samples were used to normalize the data across plexes and generate a single data matrix for downstream analysis. Results: The total number of unique proteins identified was 1,152 in experiment 1, 989 of which were measured in all samples. In experiment 2, 1,191 proteins were identified, 727 of which were measured across all samples in all plexes. Data are available via ProteomeXchange with identifier PXD025986. Spearman correlations of protein abundance estimations revealed minimal technical (0.99-1.00) and biological (0.94-0.98) variability. Each plex contained two unique pooled samples: one for normalizing across each 10plex, and one to internally validate the normalization algorithm. Spearman correlation of the validation pool following normalization was 0.86-0.90. Principal component analysis revealed stratification of samples by disease and not by plex. Subsequent differential expression and pathway analyses demonstrated significant activation of metabolic pathways and inhibition of neuroprotective pathways in proliferative diabetic retinopathy samples relative to controls.Conclusions: This study demonstrates a feasible, rigorous, and scalable method that can be applied to future proteomic studies of vitreous sampled directly from patients in order to advance understanding of pathways altered in retinal disease.

scholarly journals Correlations Between Different Angiogenic and Inflammatory Factors in Vitreous Fluid of Eyes With Proliferative Diabetic Retinopathy

2021 ◽  
Vol 8 ◽  
Author(s):  
Guanrong Wu ◽  
Baoyi Liu ◽  
Qiaowei Wu ◽  
Changting Tang ◽  
Zijing Du ◽  
...  
Keyword(s):  
Diabetic Retinopathy ◽  
Growth Factor ◽  
Proliferative Diabetic Retinopathy ◽  
Fasting Blood Glucose ◽  
Intercellular Adhesion Molecule ◽  
Inflammatory Factors ◽  
Control Group ◽  
Intercellular Adhesion Molecule 1 ◽  
Vitreous Fluid ◽  
Inflammation Mediators

Purpose: To investigate the expression of various angiogenesis and inflammation mediators in the vitreous fluid of eyes with proliferative diabetic retinopathy (PDR).Methods: A total of 38 eyes with PDR and 37 control eyes were included. Vitreous fluid was collected during vitrectomy. Vitreous levels of colony stimulating factor-1 receptor (CSF-1R), syndecan-1, placental growth factor (PIGF), and angiopoietin-like protein 4 (ANGPTL-4) were measured by multiplex immunoassay. Vitreous levels of vascular endothelial growth factor (VEGF), interleukin-6 (IL-6), interleukin-8 (IL-8), monocyte chemotactic protein-1 (MCP-1), tumor necrosis factor-α (TNF-α), and intercellular adhesion molecule-1 (ICAM-1) were measured by cytometric beads array. Levels of these mediators were compared between the PDR and control eyes. Correlations between levels of different mediators and between these mediators and kidney function metrics in the PDR group were also analyzed.Results: Vitreous levels of syndecan-1, PIGF, ANGPTL-4, VEGF, and IL-8 were significantly higher in the PDR group compared to the control group (all p < 0.05). Levels of VEGF were significantly correlated with levels of syndecan-1, PIGF, and ANGPTL-4 (r = 0.370 to 0.497, all p < 0.05). Significant positive correlations were detected between levels of any two of the following mediators including syndecan-1, PIGF, ANGPTL-4, and IL-8 (r = 0.370 to 0.906, all p < 0.05). Apart from VEGF, levels of these mediators were positively correlated with serum creatinine and blood urea nitrogen (r = 0.328 to 0.638, all p < 0.05), and negatively correlated with fasting blood glucose and estimated glomerular filtration rate (r = −0.325 to −0.603, all p < 0.05).Conclusions: Correlations between different angiogenesis and inflammation mediators were observed in eyes with PDR, suggesting cross-talks of different angiogenesis and inflammation pathways in the pathogenesis of PDR. The levels of angiogenesis and inflammation in PDR are correlated with kidney damage, indicating possible common pathways in diabetic retinopathy and nephropathy.

LIPOPOLYSACCHARIDE-BINDING PROTEIN AND SOLUBLE CD14 IN THE VITREOUS FLUID OF PATIENTS WITH PROLIFERATIVE DIABETIC RETINOPATHY

Retina ◽  
2010 ◽  
Vol 30 (2) ◽  
pp. 345-352 ◽  
Author(s):  
CRISTINA HERNÁNDEZ ◽  
FRANCISCO ORTEGA ◽  
MARTA GARCÍA-RAMÍREZ ◽  
MARTA VILLARROEL ◽  
JOAN CASADO ◽  
...  
Keyword(s):  
Diabetic Retinopathy ◽  
Proliferative Diabetic Retinopathy ◽  
Binding Protein ◽  
Lipopolysaccharide Binding Protein ◽  
Vitreous Fluid ◽  
Soluble Cd14 ◽  
Lipopolysaccharide Binding

Abstract 3764: Increased shedding of Endothelial Microparticles following Anti-VEGF therapy of Human Proliferative Diabetic Retinopathy

Circulation ◽  
2007 ◽  
Vol 116 (suppl_16) ◽  
Author(s):  
Mounir Benzerroug ◽  
Aurélie Leroyer ◽  
Serge Picaud ◽  
Alain Gaudric ◽  
Gérard Brasseur ◽  
...  
Keyword(s):  
Diabetic Retinopathy ◽  
Endothelial Cell ◽  
Intravitreal Injection ◽  
Proliferative Diabetic Retinopathy ◽  
Recombinant Antibody ◽  
Retinal Neovascularization ◽  
Complete Disappearance ◽  
Vitreous Fluid ◽  
Endothelial Microparticles ◽  
Anti Vegf

Background: Development of retinal neovascularization in proliferative diabetic retinopathy (PDR) is correlated to vitreous VEGF levels. Intravitreal administration of Bevacizumab, a humanized recombinant antibody that binds all isoforms of VEGF, causes at least short-term involution of retinal neovascularization. We hypothesized that endothelial microparticles (MP), which are submicron membrane vesicles released following endothelial cell activation or apoptosis, accumulate in vitreous fluid from patients with PDR following anti-VEGF therapy. Methods and results: Undiluted vitreous fluid samples were collected at the start of standard surgery for the treatment of retinal disease in diabetic (D, n=14, 61±3yrs, 7.5±0.2% HbA1c) and non-diabetic (ND, with macular hole, epiretinal membrane or retinal detachment; n=15, 65±4yrs) patients. Four patients with PDR received intravitreal injection of Bevacizumab (50 μL; 25 μg/ μL) one week before surgery. Levels and cellular origins of MP in vitreous fluid were analysed by flow cytometry, using markers for platelet (CD41), endothelial (CD144), microglial (Bandeiraea Simplicifolia Lectin; ILB4) and photoreceptor (Arachis hypogaea Lectin; PNA) cells. Vitreous levels of endothelial and platelet MPs were markedly increased in PDR when compared to ND patients (139±53 vs 21±5 CD41+MP/μl (p=0.02); 238±61 vs. 62±12 CD144+MP/μl (p=0.004); respectively). Levels of MPs of microglial or photoreceptor origin did not differ significantly in D and ND vitreous samples (89±51 vs. 17±6 PNA+MP/μl (p=0.234); 47±25 vs. 20±13 ILB4+MP/μl (p=0.32); respectively). Intravitreal injection of anti-VEGF antibody led to a tenfold increase in endothelial MPs shedding (2418±673 CD144+MP/μl) and a complete disappearance of platelet-derived CD41+MPs in PDR vitreous samples. Anti-VEGF treatment also reduced microglial ILB4+MPs levels (3±3 MP/μl; p<0.16). Conclusion: Microparticle identification in vitreous samples indicates that local anti-VEGF therapy induces massive vascular endothelial cell apoptosis. In addition, augmented platelet MPs levels in diabetic vitreous samples suggests that PDR is associated with an increased endothelial permeability, which is restored to basal level by anti-VEGF therapy

Sign in / Sign up

Export Citation Format

Share Document